Neurodegenerative disorders associated with protein aggregation include nine untreatable diseases caused by CAG/glutamine tract (polyQ) expansions. One of these disorders, spinobulbar muscular atrophy (SBMA), is characterized by degeneration of lower motor neurons and is caused by a mutation in the androgen receptor (AR) gene. The mutant protein undergoes hormone-dependent nuclear translocation, unfolding and oligomerization, steps that are critical to toxicity and to the development of progressive proximal limb and bulbar muscle weakness in men. Although the disease causing mutation was identified two decades ago, treatments available to SBMA patients remain largely supportive. Furthermore, the cellular pathways that degrade the mutant protein remain incompletely defined, and this lack of knowledge hinders the development of disease-modifying therapies. The objective of this application is to define the role of the Hsp90-based chaperone machinery in the protein quality control decisions that govern polyQ AR degradation. Our central hypothesis is that Hsp70 and Hsp90 have essentially opposing roles in the triage of the polyQ AR, in that Hsp70 promotes substrate ubiquitination whereas Hsp90 inhibits ubiquitination. This hypothesis springs from our preliminary data showing that association with Hsp90 stabilizes the polyQ AR, while unfolding of the mutant protein leads to ubiquitination by Hsp70-dependent E3 ligases. Here we will use genetic and pharmacological tools to define the consequences of allosterically activating Hsp70-dependent ubiquitination. Additionally, as our data point to contributions from both skeletal muscle and motor neurons to the disease phenotype, we will use genetic approaches to determine the extent to which toxicity at each site must be targeted to achieve beneficial therapeutic effects. The rationale of the proposed work is that establishing the mechanisms that regulate polyQ AR degradation will identify targets that can be exploited by the development of new therapies. Genetic and biochemical approaches will be used to determine the extent to which allosteric activators of Hsp70 promote clearance of the polyQ AR (Aim 1), to identify critical sties of polyQ AR toxicity in SBMA mice (Aim 2), and to establish the effects of novel, small molecule allosteric activators of Hsp70 in SBMA mice (Aim 3). These studies are expected to have a significant positive impact by defining pathways that limit SBMA toxicity while providing proof-of-concept data supporting a new therapeutic approach. As Hsp70 also regulates quality control decisions governing the turnover of other mutant proteins that cause neurodegeneration, we expect that the approaches defined here will inform therapeutic strategies that will be broadly applicable.